LOCUS       sequence-371122-        6831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  arabinose-inducible expression of GFP and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6831)
  AUTHORS   Avraham, Roi
  TITLE     Bacterial fluorescent reporters and tags toolkit
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6831)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pBAD_GFP_mCherry_-+
FEATURES             Location/Qualifiers
     source          1..6831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(149..862)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(189..209)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     804..832
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             871..879
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(906..1190)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1230..1249
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(1429..2307)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2334..2618
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     RBS             2645..2653
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2662..3372
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYMKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2807..2825)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3075..3094
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3603..3690
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3782..3809
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3828..3919
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3920..3988,3989..4780)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4138..4157)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4822..5277
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4909..4928)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5119..5140
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5388..5976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5877..5896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6130..6147
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6313..6335
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     terminator      complement(6452..6479)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6571..6657)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aaatcttctc tcatccgcca aaacagccaa gcttgcatgc ctgcaggtcg actctagtga
       61 taaactaccg cattaaagct tgcatgcctg caggtctgga canttttttt tttttttttt
      121 tttttttttt tttttntrym sgcctttatt tgtatagttc atccatgcca tgtgtaatcc
      181 cagcagctgt tacaaactca agaaggacca tgtggtctct cttttcgttg ggatctttcg
      241 aaagggcaga ttgtgtggac aggtaatggt tgtctggtaa aaggacaggg ccatcgccaa
      301 ttggagtatt ttgttgataa tggtctgcta gttgaacgct tccatcttca atgttgtgtc
      361 taattttgaa gttaactttg attccattct tttgtttgtc tgccatgatg tatacattgt
      421 gtgagttata gttgtattcc aatttgtgtc caagaatgtt tccatcttct ttaaaatcaa
      481 taccttttaa ctcgattcta ttaacaaggg tatcaccttc aaacttgact tcagcacgtg
      541 tcttgtagtt cccgtcatct ttgaaaaata tagttctttc ctgtacataa ccttcgggca
      601 tggcactctt gaaaaagtca tgctgtttca tatgatctgg gtatctcgca aagcattgaa
      661 caccataacc gaaagtagtg acaagtgttg gccatggaac aggtagtttt ccagtagtgc
      721 aaataaattt aagggtaagt tttccgtatg ttgcatcacc ttcaccctct ccactgacag
      781 aaaatttgtg cccattaaca tcaccatcta attcaacaag aattgggaca actccagtga
      841 aaagttcttc tcctttactc attttttata acctccttac ctcctgctag cccaaaaaaa
      901 cgggtatgga gaaacagtag agagttgcga taaaaagcgt caggtaggat ccgctaatct
      961 tatggataaa aatgctatgg catagcaaag tgtgacgccg tgcaaataat caatgtggac
     1021 ttttctgccg tgattataga cacttttgtt acgcgttttt gtcatggctt tggtcccgct
     1081 ttgttacaga atgcttttaa taagcggggt taccggtttg gttagcgaga agagccagta
     1141 aaagacgcag tgacggcaat gtctgatgca atatggacaa ttggtttctt gctcatgagc
     1201 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     1261 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat ctgctcatgt
     1321 ttgacagctt atcatcgatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca
     1381 aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt
     1441 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
     1501 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
     1561 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
     1621 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
     1681 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
     1741 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
     1801 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
     1861 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
     1921 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
     1981 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
     2041 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
     2101 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
     2161 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
     2221 gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag
     2281 caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca gagaagaaac
     2341 caattgtcca tattgcatca gacattgccg tcactgcgtc ttttactggc tcttctcgct
     2401 aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat
     2461 gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt
     2521 gcacggcgtc acactttgct atgccatagc atttttatcc ataagattag cggatcctac
     2581 ctgacgcttt ttatcgcaac tctctactgt ttctccatac ccgttttttt gggctagcag
     2641 gaggtaagga ggttataaaa aatggtgagc aagggcgagg aggataacat ggccatcatc
     2701 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     2761 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     2821 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     2881 tccaaggcct acgtgaagca ccccgccgac atccccgact acatgaagct gtccttcccc
     2941 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     3001 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     3061 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     3121 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     3181 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     3241 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     3301 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     3361 ctgtacaagt aatagagtcg acctgcaggc atgcaagctt ggctgttttg gcggatgaga
     3421 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
     3481 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
     3541 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     3601 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     3661 cggtgaacgc tctncctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     3721 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     3781 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
     3841 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3901 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3961 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4021 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4081 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4141 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     4201 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4261 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4321 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4381 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4441 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4501 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4561 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4621 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4681 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4741 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4801 atatatactt tagattgatt tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4861 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4921 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4981 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     5041 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5101 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5161 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5221 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa
     5281 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5341 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5401 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5461 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5521 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5581 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5641 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5701 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5761 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5821 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5881 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5941 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6001 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6061 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6121 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     6181 cttacgcatc tgtgcggtat ttcacaccgc atagggtcat ggctgcgccc cgacacccgc
     6241 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     6301 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     6361 cgaggcagaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     6421 cgccgaaaca agcagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg
     6481 cttaatttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg
     6541 cttcgcaacg ttcaaatccg ctcccggcgg atttgtccta ctcgggagag cgttcaccga
     6601 caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg
     6661 cctggcagtt ccctactctc gcatggggag accccacact accatcggcg ctacggcgtt
     6721 tcacttctga gttcggcatg gggtcaggtg ggaccaccgc gctactgccg ccaggcaaat
     6781 tctgttttat cagaccgctt ctgcgttctg atttaatctg tatcaggctg a
//
